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57 个结果
  • 简介:脂肪和肌肉房间,刺激胰岛素葡萄糖举起被葡萄糖transporter主要调停4(GLUT4),哪个到响应胰岛素刺激房间表面的从细胞内部分隔空间translocates。AS160Akt底层之并且调整胰岛素GLUT4translocation起重要作用。在这研究,(RUVBL2)象RuvB样蛋白质2集体spectrometry相结合哺乳动物双人脚踏车亲密关系纯化(龙头)作为新AS160有约束力蛋白质被识别。3T3-L1adipocytes,RUVBL2高度被表示并且cytosol主要是分布式adipocytesRUVBL2弄空通过减少刺激胰岛素AS160phosphorylation禁止刺激胰岛素GLUT4translocation和葡萄糖举起。然而,人RUVBL2介绍能颠倒这禁止效果。这些数据建议RUVBL2通过它和AS160相互作用在刺激胰岛素GLUT4translocation起个重要作用

  • 标签: 结合蛋白 转位 葡萄糖转运体 串联亲和纯化 脂肪细胞 肌肉细胞
  • 简介:POU抄写因素OCT4不仅在维持pluripotent和房间而且幕作为通过基因剂量个房间命运决定因素完成胚胎茎(ES)自我更新状态起个必要作用。然而,控制细胞内部OCT4蛋白质水平分子机制留下逃犯。这里,我们报导那人WWP2,E3ubiquitin(Ub)蛋白质ligase,通过它WW领域明确地OCT4交往并且vitro并且vivo提高OCT4Ub修正。我们首先证明ES房间OCT4能被Ubpost-translationally修改。而且,我们发现WWP2剂量依赖者方式,和WWP2活跃地点半胱氨酸残余通过26Sproteasome支持OCT4降级OCT4上为它活动和解朊效果被要求。显著地,我们当WWP2表示由特定RNA干扰(RNAi)downregulated时,OCT4蛋白质水平显著地被提高数据表演,建议那WWP2ES房间维持合适OCT4蛋白质水平个重要管理者。而且,北污点分析证明WWP2抄本多样的人织物/器官广泛地在场并且高度无差别的人ES房间表示。然而,它表示水平快速区分的人ES房间以后被减少,显示WWP2表示力量发展地被调整。我们调查结果证明WWP2ES房间OCT4蛋白质水平个重要管理者。

  • 标签: 人类胚胎干细胞 转录因子 退化 蛋白质水平 Northern 半胱氨酸残基
  • 简介:镁(Mg2+)充满工厂房间并且许多生理过程起关键作用。A10成员基因家庭AtMGT(也作为AtMRS2知道)Arabidopsis被识别,它属于CorA总个优核质子集,作为Mg2+transporters工作。些家庭成员(AtMGT1和AtMGT10)作为高亲密关系Mg2+transporter工作并且能补充缺乏Mg2+运输能力细菌或酵母异。这里,我们报导个AtMGT家庭成员,AtMGT9,作为低亲密关系Mg2+transporter工作,并且为花粉开发必要沙门氏菌变异紧张MM281功能互补试金证明AtMGT9能够调停在Mg2+sub-millimolar范围Mg2+举起。AtMGT9基因在成熟花药最强烈被表示并且叶子脉管纸巾,并且在年轻根也是检测。AtMGT9基因表示混乱异质接合变异+/mgt9导致成熟花粉谷物流产,并且没有同型结合变异植物selfed+/mgt9植物子孙被获得。在这些异质接合植物表示AtMGT9转基因植物能恢复花粉显型到野类型。另外,AtMGT9RNAi转基因植物也显示出类似的未成功花粉显型到变异+/mgt9起,我们结果证明AtMGT9作为男配偶体开发和男富饶起关键作用低亲密关系Mg2+transporter工作。

  • 标签: 花粉发育 拟南芥 转运 转基因植物 植物细胞
  • 简介:Mammaliancelltotipotencyisasubjectthathasfascinatedscientistsforgenerations.AlonglastingquestionwhethersomeofthesomaticcellsretainstotipotencywasansweredbythecloningofDollyattheendofthe20thcentury.Thedawnofthe218thasbroughtforwardgreatexpectationsinharnessingthepoweroftotipotentcyinmedicine.Throughstemcellbiology,itispossibletogenerateanypartsofthehumanbodybystemcellengineering.Considerableresourceswillbedevotedtoharnesstheuntappedpotentialsofstemcellsintheforeseeablefuturewhichmaytransformmedicineasweknowtoday.Atthemolecularlevel,totipotencyhasbeenlinkedtoasingulartranscriptionfactoranditsexpressionappearstodefinewhetheracellshouldbetotipotent.NamedOct4,itcanactivateorrepresstheexpressionofvariousgenes.Curiously,verylittleisknownaboutOct4beyonditsabilitytoregulategeneexpression.ThemechanismbywhichOct4specifiestotipotencyremainsentirelyunresolved.Inthisreview,wesummarizerethestructureandfunctionofOct4andaddresstoOct4functioninmaintainingtotipotencyorpluripotencyofembryonicstemcels.

  • 标签: 干细胞 全能性 复制 翻译 蛋白因子
  • 简介:InteractionbetweencytotoxicTlymphocyte-associatedantigen-4(CTLA4,CD152)andB7molecules(B7-1andB7-2)isofimportanceinthecellulareventsoflymphocyte,includingantigen-specificT-cellactivationandinductionofautoreactiveT-cell.WedescribehaerethefirstintroductionofamurinesolubleCTLA4gene,CTLA4Ig,toMm1cells,amacrophagiccellline.CTLA4IgwassuccessfullyexpressedonMm1cellsandtheexpressedCTLA4IgwasfoundtobefunctionallyactiveintheirbindingtoB7moleculesbyflowcytometryandimmunofluorescencestudies.ThebiologicalactivityofCTLA4IgfromthetransfectedMm1cellswasstudiedandshowedinhibitoryactivityonmixedlymphocyteculture.AhighCTLA4Igproducingmacrophagiccelllinewasobtained.AsMm1cellswereregardedasdifficultforgenetransfectionandtherehassofarbeennoreportonexpressionofCTLA4IggeneonMm1cells,theseresultssuggestedthattheCELA4IgexpressingMm1cellscouldbeusefulforanalysisofCTLA4andB8moleculeinteractioninbothmacrophageandT-cell.

  • 标签: T淋巴细胞相关抗原4 细胞毒 可溶型 巨噬细胞 生物学活性 表达
  • 简介:Trichosanthin(TCS)isapotentallergentomice.Accordingtoourpreviousexperiments,itcouldbringouttheIgEresponsetoovabumin(OVA)ifTCSwasgivenonedaybeforeOVAimmunization,whileOVAalonecouldnotinduceIgEtoit.Inthiswork,thekineticsofinterleukin4(IL-4)andinterferonγ(IFN-γ)geneexpressioninthemesentericlymphnode(MLN)ofTCS-immunizedmicewasinvestigatedusingasemi-quantitativeRT-PCRmethod.ItindicatedthatTCSinducedsignificantIL-4geneexpressionandthepeaksofIL4geneexpressionwereondayoneafterTCSimmunizationinbothprimaryandsecondaryresponse.Incontrast,theIFN-γgeneexpressionwassuppressed.Furthermor,theIL-4geneexpressioninthesecondaryresponsewaslowerthanthatintheprimaryresponse.ThusthepresenceofIgEmemoryBcellswerestudied.ResultsshowedthattheamountofmatureIgEmRNAarosesignificantlyandrapidlyonedayafterTCSrestimulation,whileintheMLNofthemiceprimed30daysbeforeandwithoutboost,itwasalmostasthesameamountoftheunimmunizedcontrol.ThesefindingssuggesttheexistenceoftheIgEmemoryBcellsinthemiceaftertheprimaryTCSimmunization.

  • 标签: IL-4 IFN-γ Trichosansin 小鼠 免疫反应 基因表达
  • 简介:Asimplemethodtocreateachromosome-specificDNAlibrqaryofrice,includingmicrodissection,amplification,charterizationandcloning,isdescribed.Ricechromosome4fromametaphasecellhasbeenisolatedandamplifiedbytheLinkerAdapterPCR(LA-PCR).ThePCRproductswerelabeledasprobeswithDIG-11-dUTPusingtherandomprimingmethod.SouthernblotanalysiswithricegenomicDNAandspecificRFLPmarkersdemonstratedthatthePCRproductswerederivedfromricechromosome4.Alargelibrarycomprisingover100,000recombinantplasmidmicroclonesfromricechromosome4wasconstructed.Colonyhybridizationshowedthat58%oftheclonescontainedsingleorlow-copysequencesand42%containedrepetitivesequences.ThesizeofinsertsgeneratedbyPCRrangedfrom140bpto500bp.ThismethodwillfacilitatecloningofthespecificchromosomeDNAmarkersandimportantgenesofrice.

  • 标签: 水稻 第4号染色体 DNA文库 LA-PCR 显微解剖
  • 简介:TRAF2isacriticaladaptormoleculeforTNFreceptorsininflammatoryandimmunesignaling.Uponreceptorengagement,TRAF2isrecruitedtoCD40andtranslocatestolipidraftsinaRINGfinger-dependentprocess,whichenablestheactivationofdownstreamkinases.TRAF1candisplaceTRAF2andCD40fromraftfractions,anditpromotestheabilityofTRAF2tosustainsignalactivation.ReplacementoftheRINGfingerofTRAF2witharaft-targetingsignalrestoresJNKactivationandassociationwiththecytoskeletalproteinFilamin,butnotNF-KBactivation.TRAF1-/-dendriticcellsshowattenuatedresponses

  • 标签: TRAF2细胞内定位 TRAF1调节 信号转导机制
  • 简介:Humanμ-opioidreceptor(HμOR)withatagofsixconsecutivehistidinesatitscarboxylterminushasbeenexpressedinrecombinantbaculovirusinfectedSf9insectcells.Themaximalbindingcapacityforthe[^3H]diprenorphineand[^3H]ohmefentanyl(Ohm)were9.1±0.7and6.52±0.23nmol/gprotein,respectively.The[^3H]diprenorphineor[^3H]OhmbindingtothereceptorexpressedinSf9cellswasstronglyinhibitedbyμ-selectiveagonists[D-Ala^2,N-methyl-Phe^4,glyol^5]enkephalin(DAGO),Ohm,andmorphine,butneitherbyδnorbyκselectiveagonist.Na^+(100mM)andGTP(50μM)couldreduceHμORagonistsetorphineandOhmaffinitybindingtotheoverexpressedHμOR.μ-selectiveagonistsDAGOandOhmeffectivelystimulated[^35S]GTPγSbinding(EC50=2.7nMand6.9nM)andinhibitedforskolin-stimulatedcAMPaccumulation(IC50=0.9nMand0.3nM).Theagonist-dependenteffectscouldbeblockedbyopioidantagonistnaloxoneorbypretreatmentofcellswithpertussistoxin(PTX).TheseresultsdemonstratedthatHμORoverexpressedinSf9insectcellsfunctionallycoupledtoendogenousCi/oproteins.

  • 标签: 人类μ-河片受体 Sf9昆虫细胞 过表达 百日咳毒素 内源 Gi/o蛋白
  • 简介:Interleukin-4isacytokineproducedbyactivatedTcells,mastcells,andbasophilsthatelicitsmanyimportantbiologicalresponses[1](seeTab1).TheseresponsesrangefromtheregulationofhelperTcelldifferentiation[2]andtheproductionofIgE[3]totheregulationoftheadhesivepropertiesofendothelialcellsviaVCAM-1[4],Inkeepingwiththesediversebiologicaleffects,high-affinitybindingsitesforIL-4(Kd20to300pM)havebeendetectedonmanyhematopoieticandnon-hematopoieticcelltypesatlevelsrangingfrom50to5000sitespercell[5].ThisreviewwillfocusonthediscretesignaltransductionpathwaysactivatedbytheIL-4recxeptorandthecoordinationoftheseindividualpathwaysintheregulationofafinalbiologicaloutcome.

  • 标签: 白介素-4 基因表达 生长 细胞存活 调节 生物学作用
  • 简介:<正>Usingsubtractioncloning,weidentifiedthehumanN-MycDownstream-RegulatedGene-2(hNDRG2),locatedat14q11.2,asacandidatetumorsuppressorgene.Semi-quantitativeRT-PCRshowedthattheexpressionofhNDRG2in15of27(56%)humanGBMtissuesandall6humanglioblastomacelllineswassignificantlylowerthanthatinthenormalbrain.TheexpressionofhNDRG2alsowasevaluatedin60lung-carcinomapatients.17of26casesofsquamouscarcinomaand4of11casesofsmallcelllungcancerdisplayed

  • 标签: N-Myc减量调节基因2 NDRG2 细胞生长 负向调节 癌症 表达减少
  • 简介:GelatinaseA(MMP-2)isconsideredtoplayacriticalroleincellmigrationandinvasion.Theproteinaseiscercetedfromthecellasaninactivezymogen.InvivoitispostulatedthatactivationofprogelationaseA(proMMP-2)takesplaceonthecellsurfacemediatedbymembrane-typematrixmetalloproteinases(MT-MMPs).RecentstudieshavedemonstratedthatproMMP-2isrecruitedtothecellsurfacebyinteractingwithtissueinhibitorofmetalloproteinases-2(TIMP-2)boundtoMT1-MMPbyformingaternarycomplex.FreeMT1-MMPcloselylocatedtotheternarycomplexthenactivatesproMMP-2onthecellsurface.MT1-MMPisfoundinculturedinvasivecancercellsattheinvadopodia.TheMT-MMP/TIMP-2/MMP-2systemthusprovideslocalizedexpressionofproteolysisoftheextracellularmatrixrequiredforcellmigration.

  • 标签: 胞外基质 明胶酶原 细胞表面活化 细胞迁移
  • 简介:哺乳动物胚胎,内部房间团(ICM)分离和trophectoderm(TE)房间命运选择,,被抄写因素,Oct4和Cdx2互相对抗效果调整pluripotency因素,Nanog,必要指定epiblast。我们分析Nanog和Cdx2倡导者,并且发现这二个抄写因素同样相互地被调整。用有条件TE区别的根胚胎干细胞线,我们证明Nanogoverexpression压制TE标记upregulation,当时Nanog击倒upregulatesTE标记表示。我们推进Nanog和Cdx2绑在并且镇压表演对方倡导者。而Nanog大美人在ICM导致检测Cdx2表示,我们不管多么不观察胚囊开发公开混乱,显示Nanog起到ICM和TE分离Oct4个谄媚作用

  • 标签: NANOG 哺乳动物胚胎 调控 转录因子 ICM 内细胞团
  • 简介:Perforin主要从事调停形成毛孔蛋白质目标T房间死亡并且被细胞毒素T淋巴细胞(CTL)和自然漂亮房间采用。然而,它是否也常规CD4+T房间功能起作用,仍然保持不清楚。这里,我们报导那perforin缺乏(PKO)老鼠,CD4+T房间响应Thyperproliferative房间受体(TCR)刺激。hyperproliferation这个特征被改进在房间分割并且IL-2分泌物伴随。看起来,perforin缺乏不在胸腺怒气和淋巴节点影响T房间开发。vivo,perforin缺乏导致增加抗原特定T房间增长和抗体生产。而且,PKO老鼠产生试验性自体免疫眼色素层炎。探讨分子机制,我们发现在TCR刺激以后,从PKO老鼠CD4+T房间显示增加细胞内部钙流动并且随后提高抄写因素NFAT1激活。我们结果显示perforin由影响TCR依赖Ca2+发信号调整CD4+T房间激活和免疫力反应起个否定作用

  • 标签: T细胞活化 CD4 钙信号 穿孔 细胞毒性T淋巴细胞 T细胞受体